Ortho-hydroxy-n-beta-morpholino-propiophenone fungicides



United States Patent ORTHO-HYDROXY- N-BETA-MGRPHOLINO- PROPIOPHENONEFUNGICIDES 1 Carlo Giuseppe Alherti, Alberto Vercellone, Aurelio DiMarco, and Mario Ghionepalbof Milan, Italy, assiguors to SocietaFarmaceutici Italia, Milan, Italy, a corporation of Italy No Drawing.Filed Dec. 9, 1958, SenNo. 779,075

Claims priority, application Italy Mar. 12, 1955 '4 Claims. (Cl.260-247.

This invention relates to a. new group of synthetic compounds havingpronounced fungicidal properties. The invention particularly concernsN-morpholine propionyl compounds of the general formula:

V CHr-CH; 1

aro-a-cocmcm-N 0.x

'whichX is an acid molecule moiety, and also the corresponding freebases in which X is absent. This application 'is a continuation-in-partof our a plication Serial No. 567,074, filed Feruary 23, 1956,subsequently abandoned. l: 5 I

In said general 'formula the compounds of particular present interestare those-in which A'is an'isocyclic i'a'di- -cal, such as the benzeneradical, which may be -further substituted-by halogen and alkyl,particularly iewer alkyl 7 with no more than our carbonatoms. "Nitrosubstit uents of A, and heterocyclic nuclei at, or as, A, are alsowithin the broader purview.

.,Cqm1 9u d of t is lea w i m to p se P w ful fungicides, particularlyagainst Aspergillus niger, ,3:

Candida alp i'caas, Alteljngria solani, Penicillium' notatum, Fusariumdimerum and Helmintosporium oryzae.

The process of preparing these products consists in "reacting a compoundof the general formula "with r n'orpholine hydrochloride andparaformaldehyde by v boilin g in ethanol, if necessary in the presenceof a small amount of concentrated hydrochloric acid. The para- 2,945,034fiatented July 12, 1960 ICE 2 tate obtained upon cooling is separated;it consists of 2 -hydro'iiy "4 -metiiyl s cliloropNmorpholinvpropio-'phenone hydrochloride, that, after recrystallization from methanol wafr(251) has a M.P. of 194-195 C. and

corresponds with the formula OH =1 CHFC'Hi on; -bo o'ur-'oiirri f oinoiphenonehydrochloridecrystallizes u'pon rubbing the wall formaldehyde may'be replaced by equivalent amounts of 1 aqueous formaldehyde. Theheating is extended to .1 to 10 hours; at the end of this period thereaction product crystallizes directly or after concentrating thesolution.

It is, therefore, the main object of the present inven- 'tion to providecompounds of the afore-mentioned general-formula. J

It is another object of this invention to provide a method of preparingthese new compounds.

It is still another object of the invention to employ compounds of thistype as fungicides. h

These and other objects will appear froin the following examplespresented to illustrate the invention, without the intention, however,of thereby limiting the scope of the appended claims.

- Example 1-.--3.7 gr. of -2-hydroxy-4-methyl-5-cl1loroacetophenone,dissolved in 15cc. of 95% ethanol, are refluxed for 6 hours with 2 .6 grof morpholine hydrochloride, "1.2 "gr. of paraformaldhyde and 0.1 'cc.of

concentrated hydrochloric acid. The clear reaction mixing of theunchanged starting product, and the precipirture is afilter'etl-diotfrom .t-lie' undissolved portion, consistof the reaction. .ves sel.After crystallization from meth- -anol water=( 2:1.), this compoundhasaTMf. of 197" C. It corresponds to the formula H Example foregoing'example is repeated 'With the exception that, by usingm-hydroxy-acemphenone, mhydroxy is-Nmorpholine-propiephenonehydrochloride is obtained, having a M.P. of 186-1 87 "C.,'and of theformula GET-CH2 Example 4.--5.34 gr. of p-hydroxy-acetophenone areboiled for 2 hours in 12 cc. of absolute alcohol with 550 gr. ofmorpholine hydrochloride, 1.8 -.gr. of ,para'formaldeawe and 0.1 cc. ofconcentrated hydrochloric acid =1)- Hydroxy-l'fi-N-moropholinepropiophenone hydrochloride, havingQaMP. of 2l52l7 (3., crystallizesupon cooling and-is recrystallized from methanol. It has the formulaCHPC 2 AC'H2CH2 common-rm 0.1101

G jig-03:

The following compoundszdescribed above were jected to a premliminaryscreening to establish their A supersaturated solution isobjDebaryomz/ces caajemriii.

I Debaryomyces guzllel Debaryamyces -Debrzryomyces neoformansEpidermophyton interdigitale Glenospom gruph'ii 'Hisloplasma capsulatum.

fungicidal aetivity.- The following were obtainedr teams (Table' 1compounds (in 'y/cc.) which inhibits the growth of the tested strain hasbeen checked.

strains-was determined. The results are reported in Table II whereinhibiting action is indicated by,(+), absence thereof by and (i) indioates a subinhibiting action.

TABLE H Compound Debarypmyccs marylzmdii Debar z omyces .tyrocolerGlenosporella dermatiiidis Pseudomycoderma matale Torulopsis neoformansTricophyton maggim' Tricophyton plicatilevr; Actmomyces Bostrdmi VittActinomyces Bostrdmi (NW). Actino'myces Bostrb'mi A Actmomyces BostrfimiP Nocardia asteroides 1.

For said determination the following technique was adopted: anagar-Sabouraud substrate is prepared, containing 100 'y/CC. of thetested compound. Then the molten substrate is poured in 14 cm. diameterPetri plates 6 days of incubation'at 37C., theLiLD. of the with aluminum8, hydroxyquinolate. summarized in Table IV, wherein the L.I.D.s in'y/cc. are reported, obtained in liquid substrate (yeast broth), afterTABLEI Lowest Inhibiting Dose (L. I. D.) ('y/cc.)

Compound Alter- Aapar- Penicilj i Q HeZmin- Mria gillus' Candida ZiumEusarium tospoaolam' nigger albicans notatum dimerum n'um p I oruzaeji 2Hydroxy 4 methyl 5 chloro- B N morpholino propiophenone hydrochloride Hdr g N h 1m I i h 12 100 100 100 60 so 0. y oxy- -morp o opropop enonehydrochloride (II) 12' a0 12 253- 12 12 m.Hydroxy fi N morpholinopropiophen- V onehydrochloride (III) 200 1,600v 400 400 200 200p.Hydroxy fl N morpholino propiophenone hydroel11orlde(W) 400 1 800 500800 300 200- cm. diameter plates were prepared with increasing TABLE III"dilutions of the tested materialsjin potato agar. A

suspension of spores withdrawn from an 8-days culture of the fungusstrain was deposited by streaking on the solidified agar surface; 7 1 11-As lowest inhibiting dose (L.I.D.) the lowest concentr-ation (in y/cc.)was taken, which is needed to obtain g idermo h m flocwwm 5100 runlenospora grap ii 100 00 total mhibrtlon of the growth of the fungusstrain while Saboumudms mums 10 5 keeping the plate at 28 C. durlng 5-7days. 30 Tn o hym meta m hytes- 2. 5 10 After this first screening, ourattention was particularly g l 25 centered on compounds I, H, and theinhibiting action Nocardia aatero ides 25(;l=l0) 5 ;thereof (at 100:y/cc.) on a series of pathogenlc Vfungi Compound I has been alsoexamined in comparison The results are inoculation of 8-10 days old, 1:10 diluted cultures and after 48 -96hours incubation at 38 C. p v 7 U,TABIJEIV L.I.D.-(' y/ec.

, Strains v r Al-8.hydroxy- I quinolates 48 h. 96 h 48 h. 96 h.

-Debaryomices hudeloi 50 50 50 50 Epidermophyton floccos 25 50 50 50Glenospora graphii- 25 50 50 50 Trzqophytm z radians 5 50 50 -50 25 5050 .50 5 25 50 50 Com-pound I has been also tested as an agriculturalfungicide, particularly as to its surface fungicidal activity whichproved to be outstanding. I

The term surface fungicidal activity is to define the action displayedby the compound when sprayedon the leaves or other parts of the plantswhich are to be protected. 3

For this purpose, a test method has been adopted which consists inatomizing 15 days old bean seedlings which subsequently are infected,through the'leaves, withspores of bean mildew (Uromyces'appendiculatus).

The following results have been obtained: 9

Concentration 0.03%: complete protection; no phytotoxicity phenomenon isobserved.

Concentration 0.01%: substantial decrease infection. Concentration0.005%:loss of activity. The best known fungicides for inhibition ofbean N w n4 much higher conceal-arran e:

The preparation of free Bases" eorrespondingi to' each of ave saltsdserihed" eBay-e? is aeeomptished by treatment of e correspon dinhydrochloride, or other salt thereof, wmmsrre Thisis e "pliliedfasfollowsfi 'line propiophenone hydrochloride, hereafter designated havebeenm ade In Tables I and II' the data are rep'ortedf relatingl to" thegrowth of fungi and noticed 5 and days after the sowfing. Tlie compound"C70lias proved to be abletto display, on all the tested fungi, anactivity substantially not dissimilar from the one of 'Alternari'atenuis conidia For a 4 day's aged culture on agar-carrot j are reported:for j products.

water until the 'chldro-ions have disappeared from the washing water,the product is dried at 3'5" C. under vacuum-M.P. 5456 C.

The product is the corresponding free' base, namelyortho-hydroxy-fi-N-morpholine propiophe'none, hereafter designated 0.70.The following comparative tests were made on the base. C 70 auditshydrochloride 13.18.

Rear 1%; 1

? activity of: the base C170 add ofi um albo-atrum Reinke & Berth,Alternaria tennis Nees,

. I p decoetion. has been employed; the products to be tested ,7 .a 2 1arefapplied onto slides by means of a' microsyriiigeand of 'thgforegomgt-est results It may thereonf calibrated drops of conidiasuspensionshavebeen applied; 'l l1e incubation" at 21 C. was prolonged for about 16hours.

' 'In Table 1H the percent'values of lacking germination the respectiveconcentrations of. the

Test No.- 3.=-lnflenee on the germination of conidiqof Venturia pi'r'ina (Fusicladium virescens (Bi-d.) Aderh.) agent for the pearinouldin'ess ("sp0tting) Thesampling of conidia was made directly frominfected fiearle'avesat land; the products were tested on slides as forthe'lest No. 2.

Table IYf are summarized the percent values of lacking germination ofVenturflz pi'rz'hafo'r the respective concentrations of thepredrrcts:

C 70: LD="62 p.p.m.-;LD 95:85. ppm.

Test N0. 4'.Fungicidal activity -in' vivo againstthebean rus't (Uromycesappendiculatus (Feisiflink) About 30' days aged bean plants with onlythe two primary leaves have been utilized. An aqueous sitspension. ofC70 and an aqueoussolution of B.l8 have been sprayed on the underlyingside whereon, 24 hours after treatment, the infection has been effectedwith suspension of tiredospores withdrawn from leaves of previouslymeeteepiane; Soonafter the infection, the

Endothia parasitice (Mum) Andersgnj Fusarium oxy- 5 plants have beenkept for 48 hours in a moisture Sam sporiumSchlechfl-Fusarium .solarz(Mart App. & Wz., rated room, lrthefi transferred to a green house at 26(3.; Grizphium ulmi Schwarz and Cefospof'a .beticola Sacc. mereadirrg erthe results has been made 10 days after The compound B.18 was tested inaqueous solution the infection. Finally in TableV the fungicidalactivity sterilized by filtration on Seitz Eks, whereas the com- 5'- ofboth compounds tested on bean plants against f'd CflU-w testediniacetbfiesolution} i;F or eacl 1 50 -Ur0myces appendiculates isreported, as well as the perfesetheielative controls either with wateror. acetone 'cent of disease referred to controls.

"TABLE '1' t Growth of pathogenic fungi in the presence of 6 .70 andB.18Method of mom:orated-agar'h-datii noticed am 5 days iConcentrations, p.p.m. Controls Fungi Products e 20 10 s 2 1 WaterAcetone neutmbnatnatrrcnei mm .-{g T 5* 'Veiticillium daIlZiae.-=-& {gl+++f+ '-"t eiiitiziiuni aibimhfimt gig egg g Fnsarium azyaporlum QFusanumsolani. .e.. 18 6 70 rapfilum 1mm... B18 7 liibhsjlhie rial-aw g{g Legends: --=Full inl'nb' on of growth of the tested microorganism.Full growth of the microorganism.

Growth fp th u ni f ai in he pr sen -7?) end B.18Methodofincotporated-aaar daia noticed after 1am ugg Products Once? p Controls'20 a f l '2 7 1 was Acetone Deuterobhomdtracheiphilizn;L -in gVcrticilliu'm aahzmuhiuf L- {g {g V -i- Yerticillium dZb0 -litmm. Z3 1+++++YT Aitemaria tennis Endotiu'a' ummel {g g +-h+- Fusarium oxysporium{g +'H'++ fi l Eusariunr salami"... +i'f++ Graph iu'm 1mm l ++++-i1Cercospora bezicalm. g {g PH-H- +'-l- Legends: L. .=Full inhibition ofgrowth of the tested microorganism. Eull growth of the microorganism.

TABLE III The following examples relate also to the obtainment IInfluence 0 (1.70 and B18 on theigermin atiouof Alternaria tennisconidia j Percent 0i lacking r v germination Concentration (p.p.m.) 0 7o7 B18 160. V 91 92 140. 85 77 1 79 64 100 68 45 90 58 38 8O 55 I 7 46 2560 32 9 50 25 0 4n 9 1 0 Control 11 0; '0 j v gontrolacetonen; 1 0

TABLE 1W I Influence of C.70 and B18 on the germination of Ventuiia M II a To 3.23 orure hydrochloride dissolved in 100 ml. of

of free bases from the hydrochlorides.

Exaniple 7'. Q- 5 N-morpholino -propionyl)-B-oxywater 0.9 g. NaHCOdissolved in 9. ml water are added under stirring. After 1 hour theyellow product is filtered and finally washed with distilled water; theproduct melts at 162-164 C. The product crystallizes from. methanol.

f Example 8. 2. oxy 4-methylf5 chloro-fi-N morpholino-propiophenohe aupirina conidia (fie. Fusicladium virescens). Method r v =0 the drop0n.slide.. ,w 7 I r f I a t r t r1 lair" (zoom-on N'j b r Ziiigil fifiit a Concentration(p.p.m.)

' More 31st j I -A filtered solution of-3.2 g.: hydrochloride in SOQrnl. 3g 23 water is treated under stirring andatlOC. with 0,9 g. 2?, 45NaHCO in 9 ml. water. After 1 hour the white prodi 3g uct is filteredand washed with distilled water. The prod- 22 7 55v uct melts at 91 C.,and crystallizes from aqueous 2 19 ,r methanoh83355352%;;::::::::::::::::::::::::::::::: 8 Example 9. 4- y-fip r p pTABLE v' N cm-og. Fung'icidal activity '0f'C.70 and 3.18.014 bean plantsKOO-CO G ag ainst Uromyces appendiculatus. Average dam 0 6 mow-C I.repetitions I I a. .65 ,1. g, of the hydrochloride is dissolvedon thewater bath of i j r 5 s in 25 ml. water. To the filtered solution 0.31g. NaHCO treated P left/Fred t0 g ge lg. previously dissolved in 5 ml.water are added.

leaves m 2.352 5 After said addition white bright leaflets precipitate,Percent? l which are crystallized from 4 m1. water-methanol (1:1); 070E18 070 B18 010 1518 070 B18 '70 M 1325-134? r VOtherj salts are withinthe purv ew of the invention. 0.25o0 e e 'o '0 0 0 0 0 Their preparationis obvious to chemists, for example 584 185 3.000 v I by treatment ofthe free bases with'other acids, i.e.-in-

6 6 i2n0ii do ii organic, viz. mineral, or organic acids, and by directsynthesis analogous to that Of the hydrochlorides.

min/urn 9 We claim: 7 1. As fungicidal compounds, the compounds of thegroup consisting of ortho hydroxy N-beta-morpholinopropiophenones of thefollowing formula:

R CHE-Cs,

C 0 OH2CH2N 0 R" CHr-Cg 3. As a fungicidal,ortho-hydroxy-beta-N-morpholinepropiophenone hydrochloride, of theformula I CHrCHg CLHCI CHr-C a 4. As a fungicidal, orthohydroxy-beta-N-morpholinepropiophenone, of the formula CHz-CH,

CHrC:

References Cited in the file of this patent UNITED STATES PATENTS BrusonMay 5, 1,936 ter Horst May 12, 1942 Burtner Aug. 5, 1952 Denton Aug. 23,1955 Denton Nov. 8, 1955 Schultz Jan. 22, 1957 OTHER REFERENCESTakachiyo et a1.: Jour. Pharm. Soc., Japan, vol. 76, pp. 1-3 (1956).

1. AS FUNGICIDAL COMPOUNDS, THE COMPOUNDS OF THE GROUP CONSISTING OFORTHO HYDROXY N-BETA-MORPHOLINOPROPIOPHENONES OF THE FOLLOWING FORMULA: